Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Neutralization, Flow Cytometry, CyTOF-ready

Cited:

Neutralization, Flow Cytometry, ELISA Capture

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 6401
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human TNF-alpha
Gly57-Leu233 (predicted)
Accession # P01375

Specificity

Detects human TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 25‑50% cross‑reactivity with recombinant porcine TNF‑ alpha and recombinant rhesus macaque TNF-alpha is observed but no cross-reactivity with recombinant cotton rat TNF‑ alpha, recombinant rat TNF‑ alpha, recombinant human (rh) LT alpha 1/ beta 2, rhLT alpha 2/ beta 1, rhAPRIL, rhBAFF, rhEDA‑A2, recombinant mouse EDA, rhFas Ligand, rhLIGHT, rhOX40 Ligand, rhTRAIL, rhTRANCE, rhTWEAK, or rhVEGI is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human TNF‑ alpha Antibody

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Human TNF‑ alpha Antibody.

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Human TNF‑ alpha Antibody.

Recombinant Human TNF-a (Catalog # 210-TA) induces cytotoxicity in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Human TNF-a (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Human TNF-a Monoclonal Antibody (Catalog # MAB2101). The ND50 is typically 0.015-0.06 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

Detection of TNF- alpha antibody in Human PBMC Monocytes antibody by Flow Cytometry.

Detection of TNF‑ alpha in Human PBMC Monocytes by Flow Cytometry.

Human PBMCs were treated with 1μg/ml LPS for 4 hours then stained with (A) Mouse Anti-Human TNF-a Membrane Form Monoclonal Antibody (Catalog # MAB2101) or (B) isotype control antibody (MAB002) followed by anti-Mouse IgG PE-conjugated Secondary Antibody (F0102B) and Mouse anti-Human CD14 APC-conjugated Monoclonal Antibody (FAB3832A). Staining was perfomed using our Staining Membrane-associated Proteins protocol.

Detection of Insect TNF-alpha by Western Blot

Detection of Insect TNF-alpha by Western Blot

The pre-integrated TMD docks to the cytosolic tip of the lateral gate.(A) Co-immunoprecipitation of recombinant FLAG-Sec61 alpha and Sec61 gamma. Sf21 insect cell microsomes were solubilized with 1% Deoxy BigChap (DBC) and subjected to anti-FLAG immunoprecipitation (IP). Eluted samples (Sf21) were analyzed by immunoblotting next to a sample of canine microsomal ER membranes (CRM). (B) Comparison of the relative CT7 photo-crosslinking efficiency of canine rough microsomes (CRM) and Sf21 microsomes containing wild-type FLAG-Sec61 alpha / gamma complex. CT7 crosslinking was performed as before. The relative crosslinking efficiency of recombinant Sec61 alpha was normalized to the native protein. (C) BMH crosslinking reactions using recombinant FLAG-Sec61 alpha / gamma complex. Cys49 TNF alpha 126-mers were prepared in the presence of Sf21 microsomes, and BMH crosslinking reactions were conducted as described previously. Reactions were either analyzed directly (Total) or after denaturing IP with anti-FLAG affinity resin. IP with the anti-HA affinity resin served as a specificity control. (D) Western blots of Sf21 microsomes co-expressing either wild-type or mutant FLAG-Sec61 alpha / gamma. Insect cell microsomes from cells infected with a baculovirus that lacked Sec61 genes (control virus) served as a control. (E) CT7 crosslinking of mutant Sec61 alpha / gamma complexes containing the indicated cysteine substitutions. The region of the gel corresponding to FLAG-Sec61 alpha is shown. Detection of photo-affinity labeling that is competed by CT9 indicates that the mutant Sec61 alpha / gamma complex is properly folded.DOI:https://dx.doi.org/10.7554/eLife.01483.010 Image collected and cropped by CiteAb from the following publication (https://elifesciences.org/articles/01483), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human TNF‑ alpha Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Human PBMC treated with 1 μg/ml LPS for 4 hours

Neutralization

Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. Matthews, N. and M.L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 221. The Neutralization Dose (ND50) is typically 0.015-0.06 µg/mL in the presence of 0.25 ng/mL Recombinant Human TNF‑ alpha and 1 µg/mL actinomycin D.

Flow Cytometry Panel Builder

Bio-Techne Knows Flow Cytometry

Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
Build Your Panel Now
Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-alpha, TNF- alpha, TNFA ), also known as Cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism. TNF- is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Human TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-alpha shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface. Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa molecular weight soluble trimer of the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 80 kDa TNF RII, both of which are also expressed as homotrimers present on virtually all cell types. Both type I and type II receptors bind TNF-alpha with comparable affinity, although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha.

Long Name

Tumor Necrosis Factor alpha

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

UniProt

P01375

Additional TNF-alpha Products

Product Documents for Human TNF‑ alpha Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human TNF‑ alpha Antibody

For research use only

Citations for Human TNF‑ alpha Antibody

Customer Reviews for Human TNF‑ alpha Antibody

There are currently no reviews for this product. Be the first to review Human TNF‑ alpha Antibody and earn rewards!

Have you used Human TNF‑ alpha Antibody?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

FAQs for Human TNF‑ alpha Antibody

Showing  1 - 1 的 1 FAQ Showing All

    • Q: Does Human TNF-alpha antibody, clone # 6401, detect membrane bound or soluble TNF-alpha?
      A: Human TNF-alpha antibody, Clone # 6401, binds to and neutralizes Catalog # 210-TA, which is an E. coli-derived human TNF-alpha protein, Val77-Leu233, and has been shown to be cytotoxic to L‑929 mouse fibroblast cells. According to Uniprot, the soluble form of TNF-alpha is amino acids 77-233: https://www.uniprot.org/uniprot/P01375. We have also shown binding of Clone # 6401 to the membrane bound form of TNFa via flow cytometry. We are aware of a few citations showing the antibody acts to neutralize TNFa. See https://www.rndsystems.com/products/human-tnf-alpha-antibody-6401_mab21…. Therefore, clone #6401 detects both membrane bound and soluble forms of TNF-alpha.
Showing  1 - 1 的 1 FAQ Showing All
View all FAQs for Antibodies